A radioautographic analysis of the prospective cardiac area in the chick blastoderm by means of labeled grafts

Summary Grafting regions of the blastodisc of the chick, labeled with H₃-thymidine at Stages 5 and 6 ofHamilton andHamburger, is a very useful technique for following the morphogenetic movements of the grafted material. Areas (E-M) of the blastodisc of 2.75 by 0.55 mm, grafted in the homologous region of an embryo, participate in its morphogenetic movements. The labeled reversed cardiac area can sometimes be incorporated into the host, and a tubular heart can develop which includes some of the graft tissue, Figs. 3, 4, 5, and 6. The morphogenetic movements shaping the anterior intesinal portal and early foregut can occur despite the reversal of a large rectangle of the endomesoderm, Fig. 6. The coelomic epithelium lining the operated side, a part of the myoepicardium of this side, as well as the endoderm of the ventral portion of the foregut, and, more caudally, half of the anterior intestinal portal are derived from the graft as shown by strong radioactive labeling of the cells, Figs. 3, 4 and 6. The morphogenetic movements of the graft endoderm and mesoderm are independent, the pre-heart mesoderm moving in a cranial direction, while the endoderm extends caudally, Figs. 3 and 4.This study raises an interesting question: Can the original cephalic preconal cardiogenic mesoderm regulate to form sinoatrial tissue (and vice versa) ? The evidence obtained in the present investigation is not extensive enough to warrant conclusions as to the important question of regulation; for this a considerable number of operated embryos is required in which a recognizable normal heart has developed (up to Stage 12–13).This research was supported by a grant from the Association for the Aid of Crippled Children, New York.     

Comparison of the Lipolytic Effects of Norepinephrine and BRL 37344 in Rat Brown and White Adipocytes

The lipolytic effects of norepinephrine (a non-selective β-agonist) and BRL 37344 (a selective β₃-agonist) were compared in isolated rat brown and white adipocytes. Norepinephrine and BRL 37344 maximally stimulated lipolysis in brown and white adipocytes, approximately 10 times above basal values. However, adipocyte sensitivity for BRL 37344 was greater than that for norepinephrine, particularly in brown adipocytes [the EC₅₀ values (nM) for BRL 37344 and norepinephrine were 5 ± 1 and 103 ± 31 in brown adipocytes (P <0.01) versus 56 ± 9 and 124 ± 17 in white adipocytes (P <0.05), respectively]. On the other hand, the lipolytic effects of norepinephrine were totally blocked by 20–40 times superior concentrations of propranolol or bupranolol in brown as well as in white adipocytes. In contrast, the lipolytic effects of BRL 37344 were fully inhibited by concentrations of propranolol or bupranolol that were 200–1000 superior to the β₃ agonist concentration. The results demonstrate that: (1) the (β₃-agonist BRL 37344 is as effective as norepinephrine for maximally stimulating lipolysis in rat brown and white adipocytes, (2) both adipocyte types are more sensitive to the lipolytic effects of BRL 37344 than to those of norepinephrine, (3) although bupranolol is a better antagonist than propranolol on BRL 37344-stimulated lipolysis, it cannot be considered as a specific β₃-antagonist, (4) brown adipocytes are 10 times more sensitive than white adipocytes to the lipolytic effects of BRL 37344, suggesting an important role of β₃-receptors in brown adipose tissue.     

Alkaline p-nitrophenylphosphate phosphatase activity from Halobacterium halobium. Selective activation by manganese and effect of other divalent cations.

1. Alkaline p-nitrophenylphosphate phosphatase (pNPPase) activity of Halobacterium halobium is selectively stabilized and stimulated by Mn2+ ions. 2. Mn2+ binding to native pNPPase is characterized by a dissociation constant of 0.35 mM at pH 8.5, 37 degrees C, with a Hill coefficient of 0.988. 3. Mn2+ behaves as a mixed type nonessential activator, increasing the Vmax value (beta = 6.09, pH 8.5) and decreasing the Km value for pNPP (alpha = 0.56, pH 8.5). The Ki value for inorganic phosphate (a competitive inhibitor) was also decreased in the presence of Mn2+. 4. Activation of native pNPPase by preincubation with Mn2+ is a slow temperature-dependent process, which can be described by an exponential relationship vs time. However, a weak but immediate activation was also detected. 5. Zn2+, Cu2+ and Ni2+ were found to inhibit both native and Mn(2+)-stimulated pNPPase, whereas Co2+ and Cd2+ inhibited the Mn(2+)-stimulated pNPPase but had no effect on the native enzyme form.     

Chiral Biases in Solids by Effect of Shear Gradients: A Speculation on the Deterministic Origin of Biological Homochirality

We present an experimental approach to the study of the chirality of three CM2 meteorite solid samples by direct measurement of the optical activity (circular birefringence; CB). The measurements are based on transmission two modulator generalized ellipsometry in conjuction with microscope optics to map the CB of the samples. In spite of the complexity of such optical analysis, these first results indicate the presence of optically active areas in the meteorite solid matrix. In the case of the Murchison sample the statistics of the CB mapping shows a bimodal distribution with a bias to negative CB values. The composition of the active areas probably corresponds to serpentines and other poorly identified phyllosilicate phases. The results are compatible with the hypothesis that in a mineral-based scenario for the origin of life a CB sign bias in the chiral fractures originated by mechanical and flow shear gradients on clays could be later transferred to the reactions of the absorbed organic compounds.     

The three-dimensional structure of an eukaryotic glutamine synthetase: functional implications of its oligomeric structure

The structure of the prokaryotic glutamine synthetases type I (GS-I), key enzymes in nitrogen metabolism, was determined several years ago by X-ray diffraction, and consists of a double hexameric ring. The structure of the eukaryotic GS from the plant Phaseolus vulgaris (Glutamine synthetase type II; GS-II) has now been determined at low-resolution using electron microscopy and image processing, and consists of an octamer composed of two tetramers placed back-to-back and rotated 90 degrees with respect to each other. The oligomeric structure possesses a twofold symmetry, very suggestive of each tetramer being composed of two dimers. This is reinforced by the fact that dimers are isolated as a stable albeit non-functional species during the purification procedure. Given the fact that the active site of all types of GS is formed by highly conserved residues located in the interface of two interacting monomers, the geometry of the reconstructed tetramer suggests that it only contains two functional active sites, i.e., an active site per dimer. This is supported by biochemical data, which reveal that while the octamer binds eight ATP molecules, it only binds four molecules of the transition state analogue and GS inhibitor methionine-(S)-sulfoximine-P (MetSox-P). All this suggests for the GS-II enzyme an oligomeric structure containing four active sites and four possible regulatory sites, which might point to a complex regulatory behavior.     

Differences in morphology, gas exchange and root hydraulic conductance before planting in Pinus canariensis seedlings growing under different fertilization and light regimes

As the main forestry species in the Canary Islands (Spain), Pinus canariensis is frequently used in afforestation programs. Several nursery techniques are commonly employed to modify its morphology and physiology with the aim of improving post-planting survival and growth. In this work, we studied how fertilization and light regime treatments applied during the nursery period modify biomass allocation patterns and produce effects in gas exchange and root hydraulic conductance. Seedlings were grown for a 6-month period in the nursery under two light regimes (full sunlight and 40% PAR reduction), and three fertilization levels were applied in each light regime. Morphology, biomass allocation patterns, leaf gas exchange and hydraulic conductance of the whole root system were evaluated. Fertilization treatments produced significant changes in biomass allocation, gas exchange and root hydraulic conductance under both light regimes. In contrast, no differences were found between full sunlight and shade, except for a slight variation in the root:shoot ratio. Photosynthesis rate and WUE increased with fertilization in both light regimes, while E and gs maintained the same values. An opposite trend was observed for root hydraulic conductance, which showed lower values with high fertilization regimes. The results obtained indicate that fertilization is more important, determining high photosynthetic capacity than high hydraulic conductance rates before planting in Pinus canariensis containerized seedlings.     

Characterization of SMG-9, an essential component of the nonsense-mediated mRNA decay SMG1C complex

SMG-9 is part of a protein kinase complex, SMG1C, which consists of the SMG-1 kinase, SMG-8 and SMG-9. SMG1C mediated phosphorylation of Upf1 triggers nonsense-mediated mRNA decay (NMD), a eukaryotic surveillance pathway that detects and targets for degradation mRNAs harboring premature translation termination codons. Here, we have characterized SMG-9, showing that it comprises an N-terminal 180 residue intrinsically disordered region (IDR) followed by a well-folded C-terminal domain. Both domains are required for SMG-1 binding and the integrity of the SMG1C complex, whereas the C-terminus is sufficient to interact with SMG-8. In addition, we have found that SMG-9 assembles in vivo into SMG-9:SMG-9 and, most likely, SMG-8:SMG-9 complexes that are not constituents of SMG1C. SMG-9 self-association is driven by interactions between the C-terminal domains and surprisingly, some SMG-9 oligomers are completely devoid of SMG-1 and SMG-8. We propose that SMG-9 has biological functions beyond SMG1C, as part of distinct SMG-9-containing complexes. Some of these complexes may function as intermediates potentially regulating SMG1C assembly, tuning the activity of SMG-1 with the NMD machinery. The structural malleability of IDRs could facilitate the transit of SMG-9 through several macromolecular complexes.